Myristoylated Naked2 Escorts Transforming Growth Factor α to the Basolateral Plasma Membrane of Polarized Epithelial Cells

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 101; no. 15; pp. 5571 - 5576
• Main Authors: Li, Cunxi, Franklin, Jeffrey L., Graves-Deal, Ramona, Jerome, W. Gray, Cao, Zheng, Coffey, Robert J., Scott, Matthew P.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 13-04-2004; National Acad Sciences
• Subjects: Amino Acid Motifs; Animals; Antibodies; Biological Sciences; Carrier Proteins - chemistry; Carrier Proteins - genetics; Carrier Proteins - metabolism; Cell culture techniques; Cell Line; Cell lines; Cell Membrane - metabolism; Cell membranes; Cell Polarity; Cellular biology; Codons; Complementary DNA; COS cells; Dogs; Epithelial cells; Epithelial Cells - cytology; Epithelial Cells - metabolism; Fluorescence; Immunohistochemistry; Membranes; Molecular Sequence Data; Myristic Acids - chemistry; Myristic Acids - metabolism; Plasma; Precipitin Tests; Protein Transport; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Transfection; Transforming Growth Factor alpha - metabolism; Yeasts
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.0401294101

The epidermal growth factor receptor ligands transforming growth factor α (TGFα) and amphiregulin are delivered to the basolateral surface of polarized epithelial cells where they are cleaved by TACE/ADAM17. Basolateral sorting information resides in their cytoplasmic tail domains, but tail-interacting proteins required for basolateral trafficking have not been identified. Naked (NKD)1 and NKD2 are mammalian homologs of Drosophila Naked Cuticle, which negatively regulates canonical Wnt signaling by binding Dishevelled. We present evidence that NKD2, but not NKD1, binds to basolateral sorting motifs in the cytoplasmic tail of TGFα. Processing and cell-surface delivery of TGFα are accelerated in NKD2-overexpressing Madin-Darby canine kidney cells. NKD2 is myristoylated on glycine, the second residue. On expression of myristoylation-defective (G2A) NKD2, neither NKD2 nor TGFα appears at the basolateral plasma membrane of polarized Madin-Darby canine kidney cells; however, membrane staining for TGFα is restored on silencing expression of this mutant NKD2. Amphiregulin does not interact with NKD2 and retains its basolateral localization in G2A-NKD2-expressing cells, as do Na+, K+ ATPase α1 and E-cadherin. These data identify an unexpected function for NKD2, i.e., myristoylation-dependent escort of TGFα to the basolateral plasma membrane of polarized epithelial cells.